Cyclopentan-2-ol-1-yl-[1,2,3]triazolo[4,5-D]pyrimidine compounds

ABSTRACT

The invention provides novel 1,2,3-triazolo[4,5-d]pyrimidine compounds, such as those represented by formula (I):
 
                 
 
their use as medicaments, particularly in platelet aggregation disorders, compositions containing them and processes for their preparation.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a national stage filing under 35 U.S.C. 371 ofInternational Application No. PCT/SE01/02471, filed Nov. 7, 2001, whichclaims priority from Sweden Patent Application No. 0004099-8, filed Nov.9, 2000, the specifications of each of which are incorporated byreference herein. International Application No. PCT/SE01/02471 waspublished under PCT Article 21 (2) in English.

FIELD OF THE INVENTION

The present invention provides novel 1,2,3-triazolo[4,5-d]pyrimidineanalogues, their use as medicaments, compositions containing them andprocesses for their preparation.

BACKGROUND OF THE INVENTION

Platelet adhesion and aggregation are initiating events in arterialthrombosis. Although the process of platelet adhesion to thesub-endothelial surface may have an important role to play in the repairof damaged vessel walls, the platelet aggregation that this initiatescan precipitate acute thrombotic occlusion of vital vascular beds,leading to events with high morbidity such as myocardial infarction andunstable angina. The success of interventions used to prevent oralleviate these conditions, such as thrombolysis and platelet-mediatedocclusion or re-occlusion also compromises angioplasty.

A number of converging pathways lead to platelet aggregation. Whateverthe initial stimulus, the final common event is a cross-linking ofplatelets by binding of fibrinogen to a membrane-binding site,glycoprotein IIb/IIIa (GPIIb/IIIa). The high anti-platelet efficacy ofantibodies or antagonists for GPIIb/IIIa is explained by theirinterference with this final common event. However, this efficacy mayalso explain the bleeding problems that have been observed with thisclass of agent. Thrombin can produce platelet aggregation largelyindependently of other pathways but substantial quantities of thrombinare unlikely to be present without prior activation of platelets byother mechanisms. Thrombin inhibitors such as hirudin are highlyeffective anti-thrombotic agents, but again may produce excessivebleeding because they function as both anti-platelet and anti-coagulantagents. (The TIMI 9a Investigators (1994), Circulation 90, pp.1624–1630; The Global Use of Strategies to Open Occluded CoronaryArteries (GUSTO) IIa Investigators (1994) Circulation 90, pp. 1631–1637;Neuhaus K. L. et. al. (1994) Circulation 90, pp. 1638–1642.)

It has been found that ADP acts as a key mediator of thrombosis. Apivotal role for ADP is supported by the fact that other agents, such asadrenaline and 5-hydroxytryptamine (5HT, serotonin) will only produceaggregation in the presence of ADP. The limited anti-thrombotic efficacyof aspirin may reflect the fact that it blocks only one source of ADPwhich is that released in a thromboxane-dependent manner followingplatelet adhesion (see e.g. Antiplatelet Trialists' Collaboration(1994), Br. Med. J. 308, pp. 81–106 and Antiplatelet Trialists'Collaboration (1994), Br. Med. J. 308, pp. 159–168). Aspirin has noeffect on aggregation produced by other sources of ADP, such as damagedcells or ADP released under conditions of turbulent blood flow.

ADP-induced platelet aggregation is mediated by the P_(2T) receptorsubtype located on the platelet membrane. The P_(2T) receptor (alsoknown as P2Y_(ADP) or P2T_(AC)) is primarily involved in mediatingplatelet aggregation/activation and is a G-protein coupled receptor,which is as yet uncloned. The pharmacological characteristics of thisreceptor have been described, for example, in the references byHumphries et al., Br. J. Pharmacology, (1994), 113, 1057–1063, andFagura et al., Br. J. Pharmacology (1998) 124, 157–164. Recently it hasbeen shown that antagonists at this receptor offer significantimprovements over other anti-thrombotic agents (see J. Med. Chem. (1999)42, 213). Accordingly there is a need to find further P_(2T) (P2Y_(ADP)or P2T_(AC)) antagonists as anti-thrombotic agents.

DESCRIPTION OF THE INVENTION

In a first aspect the invention therefore provides a compound of formula(I):

wherein:

-   R¹ is hydrogen or hydroxy;-   R₂ is C₁₋₆ alkyl, or C₁₋₆ haloalkyl, or phenyl optionally    substituted by halogen or by C₁₋₆ alkyl, optionally substituted by    halogen;-   R³ is C₃₋₆ cycloalkyl, optionally substituted by R⁴;-   R⁴ is hydrogen or phenyl, optionally substituted by alkyl C₁₋₆,    halogen, or C₁₋₆ alkoxy.

Preferably the compound of formula (I) has the followingstereochemistry:

where R¹, R² and R³ and are as defined above.

When R³ is

where R⁴ is defined above, the stereochemistry is preferably

-   Suitably, R¹ is hydrogen or hydroxy;-   Suitably, R² is C₁₋₄ alkyl, or C,₁₋₄ haloalkyl, or phenyl optionally    substituted by halogen or by C₁₋₄ alkyl, optionally substituted by    halogen;-   Suitably, R³ is cyclopropyl, optionally substituted by R⁴;-   Suitably, R⁴ is hydrogen or phenyl, optionally substituted by    halogen or C₁₋₄ alkoxy.

Particularly preferred compounds of the invention include:

-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Methylphenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(methylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(ethylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(3,4-dichlorophenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1    ,2-diol;-   [1R-[1α,2α,3β)]]-3-[7-(Cyclopropylamino)-5-(4-trifluoromethylphenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(3,3,3,-trifluoropropylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Chlorophenyl)cyclopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Fluorophenyl)cyclopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;-   [(1R-[1α,2β(1R*,2S*)]]-2-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol;-   [1R-[1α,2β(1R*,25*)]]-2-[[7-[2-(4-Methoxyphenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol;-   [1R-[1α,2β(1R*,2S*)]]-2-[7-[[2-(3,4-Difluorophenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol;    or a pharmaceutically acceptable salt or solvate thereof, or a    solvate of such a salt.

Compounds of formula (I) can be prepared by:

-   a) The dihydroxylation of a compound of formula (II),

where R² and R³ are as defined above, preferably using osmium tetroxide,in the presence of an oxidising agent, preferablyN-methylmorpholine-N-oxide, under aqueous conditions, preferably inaqueous tetrahydrofuran, preferably at a temperature between 20° C. and50° C.

Compounds of formula (II), where R² and R³ are as defined above, may beprepared by reaction of a compound of formula (III),

where R² and R³ are as defined above, by conversion of the hydroxylgroup to an ester by treatment with an acylating agent, preferablyacetyl chloride, in an inert solvent, preferably dichloromethane, in thepresence of a base, preferably pyridine or 4-dimethylaminopyridine, at atemperature between 10° C. and 50° C. and then reductive removal of theester function by treatment with a reducing agent, preferably sodiumborohydride, in the presence of a Pd(0) catalyst, preferablytetrakis(triphenylphosphine)palladium(0), in an inert solvent,preferably tetrahydrofuran or a hindered alcohol.

Compounds of formula (III), where R² and R³ are as defined above, may beprepared by reaction of a compound of formula (IV),

where R² is defined above, with a compound of formula R³NH₂, where R³ isas defined above, in the presence of a base, preferablyN,N-di-isopropylethylamine, in an inert solvent, preferablytetrahydrofuran or dichloromethane, at a temperature between 10° C. and50° C.

Compounds of formula (IV), where R² is defined above, may be prepared byreaction of a compound of formula (V),

where R² is defined above, with 4-amino-2-cyclopenten-1-ol in an inertsolvent, preferably tetrahydrofuran, in the presence of a base,preferably triethylamine, at a temperature between 10° C. and 50° C.,and then reduction of the nitro group by treatment with an appropriatereducing agent, preferably a suspension of iron powder in an acidicsolvent, preferably acetic acid, at a temperature between 10° C. and 50°C., followed by closure of the triazole ring by the use of a nitrosatingagent, preferably iso-amyl nitrite, in an inert solvent, preferablyacetonitrile, at a temperature between 20° C. and 90° C.

-   b) The reaction of a compound of formula (VI),

where R¹ and R² are as defined above, with a compound of formula R³NH₂,eg a compound of formula (VII),

where R⁴ is as defined above, in the presence of a base, preferablyN,N-di-isopropylethylamine, in an inert ethereal solvent, preferablydiethyl ether or tetrahydrofuran or a chlorocarbon solvent, preferablydichloromethane, at a temperature between 20° C. and 50° C.

Where R⁴ is phenyl, (1R-trans)-2-phenylcyclopropanamine,[R-(R*,R*)]-2,3-dihydroxybutanedioate (1:1), the compound of formula(IV) may be prepared as described by L. A. Mitscher et al, J. Med.Chem., 1986, 29, 2044. Where R⁴ is substituted phenyl, the compound offormula (VII) may be prepared as described in WO 9905143.

Compounds of formula (VI), where R¹ and R² are as defined above, may beprepared by reaction of a compound of formula (V), where R² is definedabove, with a compound of formula (VIII),

where R¹ is as defined above, in an inert solvent, preferablytetrahydrofuran, in the presence of a base, preferably triethylamine, ata temperature between 10° C. and 50° C., and reduction of the nitrogroup by treatment with an appropriate reducing agent, preferably asuspension of iron powder in an acidic solvent preferably acetic acid,at a temperature between 10° C. and 50° C., followed by closure of thetriazole ring by the use of a nitrosating agent, preferably iso-amylnitrite, in an inert solvent, preferably acetonitrile, at a temperaturebetween 20° C. and 90° C.

-   c) The reaction of a compound of formula I, where R¹ and R² are as    defined above, with an oxidising agent, preferably    3-chloroperoxybenzoic acid, in an inert solvent, preferably    dichloromethane, at a temperature between 10° C. and 50° C.,    followed by reaction of the thus formed sulphonyl compound with a    compound of formula R^(2′)SM, where R^(2′) is a different group R²    as defined above and M is a group I or II metal, preferably sodium,    in an inert solvent, preferably tetrahydrofuran, at a temperature    between 10° C. and 50° C.

All novel intermediates form a further aspect of the invention.

Salts of the compounds of formula (I) may be formed by reacting the freebase, or a salt or a derivative thereof, with one or more equivalents ofthe appropriate acid (for example a hydrohalic (especially HCl),sulphuric, oxalic or phosphoric acid). The reaction may be carried outin a solvent or medium in which the salt is insoluble or in a solvent inwhich the salt is soluble, e.g. water, ethanol, tetrahydrofuran ordiethyl ether, which may be removed in vacuo, or by freeze drying. Thereaction may also be a metathetical process or it may be carried out onan ion exchange resin. The non-toxic physiologically acceptable saltsare preferred, although other salts may be useful, e.g. in isolating orpurifying the product.

The compounds of the invention act as P_(2T) (P2Y_(ADP) or P2T_(AC))receptor antagonists. Accordingly, the compounds are useful in therapy,including combination therapy, particularly they are indicated for useas: inhibitors of platelet activation, aggregation and degranulation,promoters of platelet disaggregation, anti-thrombotic agents or in thetreatment or prophylaxis of unstable angina, coronary revascularisationprocedures including angioplasty (PTCA), myocardial infarction,perithrombolysis, primary arterial thrombotic complications ofatherosclerosis such as thrombotic or embolic stroke, transientischaemic attacks, peripheral vascular disease, myocardial infarctionwith or without thrombolysis, arterial complications due tointerventions in atherosclerotic disease such as angioplasty,endarterectomy, stent placement, coronary and other vascular graftsurgery, thrombotic complications of surgical or mechanical damage suchas tissue salvage following accidental or surgical trauma,reconstructive surgery including skin and muscle flaps, conditions witha diffuse thrombotic/platelet consumption component such as disseminatedintravascular coagulation, thrombotic thrombocytopaenic purpura,haemolytic uraemic syndrome, thrombotic complications of septicaemia,adult respiratory distress syndrome, anti-phospholipid syndrome,heparin-induced thrombocytopaenia and pre-eclampsia/eclampsia, or venousthrombosis such as deep vein thrombosis, venoocclusive disease,haematological conditions such as myeloproliferative disease, includingthrombocythaemia, sickle cell disease; or in the prevention ofmechanically-induced platelet activation in vivo, such ascardio-pulmonary bypass and extracorporeal membrane oxygenation(prevention of microthromboembolism), mechanically-induced plateletactivation in vitro, such as use in the preservation of blood products,e.g. platelet concentrates, or shunt occlusion such as in renal dialysisand plasmapheresis, thrombosis secondary to vascular damage/inflammationsuch as vasculitis, arteritis, glomerulonephritis, inflammatory boweldisease and organ graft rejection, conditions such as migraine,Raynaud's phenomenon, conditions in which platelets can contribute tothe underlying inflammatory disease process in the vascular wall such asatheromatous plaque formation/progression, stenosis/restenosis and inother inflammatory conditions such as asthma, in which platelets andplatelet-derived factors are implicated in the immunological diseaseprocess. Further indications include treatment of CNS disorders andprevention of the growth and spread of tumours.

According to the invention there is further provided the use of acompound according to the invention as an active ingredient in themanufacture of a medicament for use in the treatment or prevention ofthe above disorders. In particular the compounds of the invention areuseful for treating myocardial infarction, thrombotic stroke, transientischaemic attacks, peripheral vascular disease and stable and unstableangina, especially unstable angina. The invention also provides a methodof treatment or prevention of the above disorders which comprisesadministering a therapeutically effective amount of a compound accordingto the invention to a person suffering from or susceptible to such adisorder.

The compounds may be administered topically, e.g. to the lung and/or theairways, in the form of solutions, suspensions, HFA aerosols and drypowder formulations; or systemically, e.g. by oral administration in theform of tablets, pills, capsules, syrups, powders or granules, or byparenteral administration in the form of sterile parenteral solutions orsuspensions, by subcutaneous administration, or by rectal administrationin the form of suppositories or transdermally.

The compounds of the invention may be administered on their own or as apharmaceutical composition comprising the compound of the invention incombination with a pharmaceutically acceptable diluent, adjuvant orcarrier. Particularly preferred are compositions not containing materialcapable of causing an adverse, e.g. an allergic, reaction.

Dry powder formulations and pressurised HFA aerosols of the compounds ofthe invention may be administered by oral or nasal inhalation. Forinhalation the compound is desirably finely divided. The compounds ofthe invention may also be administered by means of a dry powder inhaler.The inhaler may be a single or a multi dose inhaler, and may be a breathactuated dry powder inhaler.

One possibility is to mix the finely divided compound with a carriersubstance, e.g. a mono-, di- or polysaccharide, a sugar alcohol oranother polyol. Suitable carriers include sugars and starch.Alternatively the finely divided compound may be coated by anothersubstance. The powder mixture may also be dispensed into hard gelatinecapsules, each containing the desired dose of the active compound.

Another possibility is to process the finely divided powder intospheres, which break up during the inhalation procedure. Thisspheronized powder may be filled into the drug reservoir of a multidoseinhaler, e.g. that known as the Turbuhaler® in which a dosing unitmeters the desired dose which is then inhaled by the patient. With thissystem the active compound with or without a carrier substance isdelivered to the patient.

The pharmaceutical composition comprising the compound of the inventionmay conveniently be tablets, pills, capsules, syrups, powders orgranules for oral administration; sterile parenteral or subcutaneoussolutions, suspensions for parenteral administration or suppositoriesfor rectal administration.

For oral administration the active compound may be admixed with anadjuvant or a carrier, e.g. lactose, saccharose, sorbitol, mannitol,starches such as potato starch, corn starch or amylopectin, cellulosederivatives, a binder such as gelatine or polyvinylpyrrolidone, and alubricant such as magnesium stearate, calcium stearate, polyethyleneglycol, waxes, paraffin, and the like, and then compressed into tablets.If coated tablets are required, the cores, prepared as described above,may be coated with a concentrated sugar solution, which may contain e.g.gum arabic, gelatine, talcum, titanium dioxide, and the like.Alternatively, the tablet may be coated with a suitable polymerdissolved either in a readily volatile organic solvent or an aqueoussolvent.

For the preparation of soft gelatine capsules, the compound may beadmixed with e.g. a vegetable oil or polyethylene glycol. Hard gelatinecapsules may contain granules of the compound using either the abovementioned excipients for tablets, e.g. lactose, saccharose, sorbitol,mannitol, starches, cellulose derivatives or gelatine. Also liquid orsemisolid formulations of the drug may be filled into hard gelatinecapsules.

Liquid preparations for oral application may be in the form of syrups orsuspensions, for example solutions containing the compound, the balancebeing sugar and a mixture of ethanol, water, glycerol and propyleneglycol. Optionally such liquid preparations may contain colouringagents, flavouring agents, saccharine and carboxymethylcellulose as athickening agent or other excipients known to those skilled in art.

EXAMPLES

The invention is illustrated by the following non-limiting examples.

In the examples the NMR spectra were measured on a Varian Unity Inova300 or 400 spectrometer and the MS spectra were measured as follows: EIspectra were obtained on a VG 70-250S or Finnigan Mat Incos-XLspectrometer, FAB spectra were obtained on a VG70-250SEQ spectrometer,ESI and APCI spectra were obtained on Finnigan Mat SSQ7000 or aMicromass Platform spectrometer. Preparative HPLC separations weregenerally performed using a Novapak®, Bondapak® or Hypersil® columnpacked with BDSC-18 reverse phase silica. Flash chromatography(indicated in the Examples as (SiO₂)) was carried out using FisherMatrix silica, 35–70 μm. For examples which showed the presence ofrotamers in the proton NMR spectra only the chemical shifts of the majorrotamer are quoted.

Example 1[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a)(1S-cis)-4-[[6-Chloro-5-nitro-2-(propylthio)-pyrimidin-4-yl]amino]-2-cyclopenten-1-ol

To a solution of 4,6-dichloro-5-nitro-3-propylthiopyrimidine (preparedas described in WO 9703084) (4.00 g) and triethylamine (2.00 ml) in drytetrahydrofuran (100 ml) was added dropwise over 1 hour a solution of[1S-cis]-4-amino-2-cyclopenten-1-ol (prepared as described by S. F.Martin et al., Tetrahedron Lett., 1992, 33, 3583) (1.48 g) in a mixtureof tetrahydrofuran (100 ml) and 1,4-dioxane (50 ml). The reactionmixture was filtered, concentrated in vacuo and the residue purified bychromatography (SiO₂, ethyl acetate:isohexane 1:4 to 1:1 as eluant) toafford the sub-title compound (3.18 g).

MS (APCI) 313 (M−H₂O+H⁺, 100%).

b)(1S-cis)-4-[[5-Amino-6-chloro-2-(propylthio)pyrimidin-4-yl]amino]-2-cyclopenten-1-ol

Iron powder (2.30 g) was added to a stirred solution of the product ofstep a) (2.61 g) in acetic acid (100 ml). The reaction mixture wasstirred at room temperature for 2 hours, concentrated in vacuo to halfvolume, diluted with ethyl acetate and washed with water. The organiclayer was dried and concentrated in vacuo to afford the sub-titlecompound (2.28 g).

NMR δH (d₆-DMSO) 7.03 (1H,d), 5.93–5.90 (1H, m), 5.85–5.82 (1H, m),5.05(1H, d), 4.91–4.85 (2H, m), 4.66–4.60 (1H, m), 2.94 (2H, t),2.77–2.68 (1H, m), 1.69–1.57 (2H, sextuplet), 1.48–1.42 (1H,quintuplet), 0.94 (3H, t).

c)(1S-cis)-4-[7-Chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]-2-cyclopenten-1-ol

Isoamyl nitrite (1.08 ml) was added to a solution of the product of stepb) (2.20 g) in acetonitrile (100 ml) and the solution heated at 70° C.for 1 hour. The cooled reaction mixture was concentrated in vacuo andthe residue purified by chromatography (SiO₂, ethyl acetate:isohexane1:2 as eluant) to afford the subtitle compound (1.79 g).

MS (APCI) 312 (M+H⁺), 224 (100%).

d)[1S-[1α,4α(1S*,2R*)]]-4-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]-2-cyclopenten-1-ol

A solution of the product from step (c) (0.65 g),(1R-trans)-2-phenyl-cyclopropanamine,[R-(R*,R*)]-2,3-dihydroxybutanedioate (1:1) (prepared as described by L.A. Mitscher et al., J. Med. Chem. 1986, 29, 2044) (0.65 g) andN,N-diisopropylethylamine (1.1 ml) in dichloromethane (20 ml) wasstirred at room temperature for 16 hours. The reaction mixture wasconcentrated in vacuo and the residue purified by chromatography (SiO₂,ethyl acetate:hexane 1:2 as eluant) to afford the sub-title compound(0.786 g).

MS (APCI) 409 (M+H⁺, 100%)

e)[1S-[1α,4α(1S*,2R*)]]-4-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]-2-cyclopentene-1-ol, acetate (ester)

To a solution of the product of step d) (0.78 g), pyridine (0.43 ml) and4-dimethylaminopyridine (1 mg) in dichloromethane (15 ml) was addedacetyl chloride (0.16 ml). The solution was stirred for 3 hours and thenconcentrated in vacuo. The residue was purified by chromatography (SiO₂,ethyl acetate:hexane 1:1 as eluant) to afford the sub-title compound(0.75 g).

MS (APCI) 451 (M+H⁺, 100%).

f)[1R-[1α(1R*,2S*)]]-3-(2-Cyclopenten-1-yl)-N-(2-phenylcyclopropyl)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-7-amine(and(1R-trans)-3-(3-cyclopenten-1-yl)-N-(2-phenylcyclopropyl)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-7-amine)

To a mixture of the product from step e) (0.50 g) and sodium borohydride(0.21 g) in tetrahydrofuran (10 ml) and 2-propanol (10 ml) was addedtetrakis(triphenylphosphine)palladium(0) (6 mg). The reaction mixturewas stirred for 10 minutes, concentrated in vacuo and the residuepurified by chromatography (SiO₂, ethyl acetate:hexane 1:10 as eluant)to afford the sub-title compounds (0.38 g) as a 3:1 mixture that wasused without further purification.

MS (APCI) 393 (M+H⁺, 100%)

g)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

To a mixture of the products from step f) (0.34 g),N-methylmorpholine-N-oxide (0.20 g), tetrahydrofuran (10 ml) and water(1 ml) was added osmium tetroxide (0.9 ml, 2.5% solution in t-butanol).The mixture was stirred at room temperature for 16 hours and thentreated with sodium hydrosulphite (0.15 g) and water (1 ml). Thesuspension was filtered through Celite and the filtrate concentrated invacuo. The residue was purified by chromatography (SiO₂, ethylacetate:hexane 1:1 as eluant) to afford the title compound (0.14 g).

MS (APCI) 427 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 0.82 and 0.99 (3H, t), 1.21–1.76 (5H, m), 1.95–2.37(4H, m), 2.80–3.10 (2H, m), 3.18–3.21 and 3.82–3.87 (1H, m), 4.04 (1H,m), 4.46 (1H, m), 4.76 (1H, d), 5.01–5.05 (2H, m), 7.16–7.21 (3H, m),7.27–7.31 (2H, m), 9.33 (1H, d).

Example 2[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Methylphenyl)cyclopropyl]amino]5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a) (3R)-3-[Bis[(1,1-dimethylethoxy)carbonyl]amino]-1-cyclopentene

To a solution of(1S-cis)-4-(di-tert-butoxycarbonylamino)cyclopent-2-enyl acetate(prepared as described by D. Zhang et.al. Tett. Lett. 1996, 3799–3802)(8.98 g), tetrakis(triphenylphosphine)palladium(0) (0.463 g) inisopropanol (135 ml) and tetrahydrofuran (135 ml) was added sodiumborohydride (4.97 g) portionwise at 0° C. The mixture was stirred at 0°C. for 3 hours before the careful dropwise addition of glacial aceticacid (20 ml). The solution was concentrated in vacuo and the residuepurified by chromatography (SiO₂, diethylether 1:9 isohexane as eluent)to afford the sub-title compound (5.90 g).

NMR δH (CDCl₃) 5.82 (1H, m), 5.70–5.61 (2H, m), 5.20 (1H, m), 2.65–1.85(3H, m), 1.49–1.48 (18H, s).

b)[1R-(1α,2α,3β)]-1-[Bis[(1,1-dimethylethoxy)carbonyl]amino]-2,3-dihydroxycyclopentane

To a mixture of the product from step a) (5.90 g),N-methylmorpholine-N-oxide (3.19 g), tetrahydrofuran (150 ml) and water(15 ml) was added osmium tetraoxide (10.60 ml, 2.5% solution int-butanol). The mixture was stirred at room temperature for 16 hours andthen treated with sodium hydrosulphite (3.50 g) and water (50 ml). Thesuspension was filtered through Celite and the filtrate concentrated invacuo. The residue was purified by chromatography (SiO₂, ethyl acetate:hexane 1:2 as eluant) to afford the sub-title compound (4.135 g).

NMR δH (d₆-DMSO) 4.55–4.41 (2H, m), 4.34–4.08 (2H, m), 3.83–3.81 (1H,m), 1.90–1.47 (4H, m), 1.44 (18H, s).

c) [1R-(1α,2α,3β)]-3-Aminocyclopentane-1,2-diol, hydrochloride

To a solution of product from step b) (4.1 g) in methanol (40 ml) wasadded conc. hydrochloric acid (10 ml). The solution was stirred for 4hours and concentrated in vacuo to give an oil, which was azeotropedwith toluene to afford the sub-title compound (3.10 g).

NMR δH (d₆-DMSO) 8.28 (2H, s), 4.63 (3H, s), 3.91–3.78 (2H, m),3.26–3.20 (1H, m), 2.10–1.80 (2H, m), 1.52–1.49 (2H, m).

d)[1R-(1α,2α,3β)]-3-[[6-Chloro-5-nitro-2-(propylthio)-pyrimidin-4-yl]amino]cyclopentane-1,2-diol

A solution of the product from step c) (3.10 g) in dry tetrahydrofuran(100 ml) was added dropwise over 1 hour to a solution of4,6-dichloro-5-nitro-3-propylthiopyrimidine (prepared as described in WO9703084) (7.00 g) and N,N-diisopropylethylamine (11.30 ml) in drytetrahydrofuran (100 ml). The reaction mixture was heated to reflux for20 hours and concentrated in vacuo. The residue was purified bychromatography (SiO₂, ethyl acetate:isohexane 1:1 as eluant) to affordthe sub-title compound (3.79 g)

NMR δH (CDCl₃) 7.97–7.95 (1H, d), 4.48–4.43 (1H, m), 4.18–4.08 (2H, d),3.94–3.91 (1H, m), 3.18–3.08 (2H, m), 2.50–2.43 (1H, m), 2.09–2.07 (1H,m), 1.90–1.87 (1H, m), 1.81–1.73 (2H, q), 1.60–1:54 (2H, m), 1.08–1.03(3H, t).

e)[1R-(1α,2α,3β)]-3-[[5-Amino-6-chloro-2-(propylthio)-pyrimidin-4-yl]amino]cyclopentane-1,2-diol

Iron powder (3.80 g) was added to a stirred solution of the product ofstep d) (3.80 g) in acetic acid (50 ml). The reaction mixture wasstirred at room temperature for 2 hours, concentrated in vacuo to halfvolume, diluted with ethyl acetate and washed with water. The organiclayer was dried and concentrated in vacuo to afford the sub-titlecompound (3.36 g).

MS (APCI) 319 (M+H⁺) (100%).

f)[1R-(1α,2α,3β)]-3-[7-Chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

A solution of sodium nitrite (1.23 g) in water (5 ml) was added dropwiseto solution of product from step e) (3.36 g) in acetic acid (50 ml). Thereaction mixture was stirred at room temperature for 2 hours andconcentrated in vacuo. The residue purified by chromatography (SiO₂,ethyl acetate:isohexane 1:1 as eluant) to afford the sub-title compound(2.20 g).

MS (APCI) 302 (M+H⁺) loss of N₂ (100%).

NMR δH (CDCl₃) 5.27–5.18 (1H, m), 4.69–4.65 (1H, m), 4.42–4.38 (1H, m),3.21–3.16 (2H, t), 2.69–2.59 (1H, m), 2.39–2.26 (2H, m), 2.09–1.98 (1H,m), 1.87–1.75 (2H, m), 1.11–1.06 (3H, t).

g)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Methylphenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

A solution of the product from step f) (0.150 g),(1R-trans)-2-(4-methylphenyl)cyclopropanamine,[R-(R*,R*)]-2,3-dihydroxybutanedioate (1:1) (prepared as described WO9905143) (0.65 g) and N,N-diisopropylethylamine (0.155 ml) in dioxane(20 ml) was stirred at room temperature for 16 hours. The reactionmixture was concentrated in vacuo and the residue purified bychromatography (SiO₂, ethyl acetate:dichloromethane 1:2 as eluant) toafford the sub-title compound (0.120 g).

MS (APCI) 441 (M+H^(+, 100)%)

NMH δH (d₆-DMSO) 9.30–9.29 (1H, d), 7.08–7.05 (4H, s), 5.06–4.75 (3H,m), 4.49–4.42 (1H, m), 4.05 (1H, m), 3.16–2.86 (3H, m), 2.26 (3H, s),2.12–1.97 (3H, m), 1.73–1.26 (3H, m), 0.85–0.80 (3H, t)

Example 3[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[-2-Phenylcyclopropyl)amino]-5-(methylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared as described in Example 1, step d) using(1R-trans)-2-phenylcyclopropanamine and the product from Example 2, stepf).

MS (APCI) 427 (M+H⁺, 100%).

b)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(propylsulphonyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

A solution of product from step a) (1.50 g) in dichloromethane wastreated with 3-chloroperoxybenzoic acid (2.42 g). The solution wasstirred at room temperature for 2 hours before washing organic layerwith sodium metabisulphite solution. The organic layer was dried (MgSO₄)and concentrated in vacuo. The residue was purified by chromatography(SiO₂, ethyl acetate as eluant) to afford the sub-title compound (1.30g).

MS (APCI) 459 (M+H⁺, 100%).

c)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(methylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

A solution of product from step b) (0.15 g) in tetrahydrofuran (10 ml)was treated with sodium methanethiolate (0.046 g) in water (1 ml). Thesolution was stirred at room temperature overnight. The solution wasconcentrated in vacuo and residue was purified by chromatography (SiO₂,ethyl acetate:dichloromethane 1:1 as eluant) to afford the titlecompound (0.095 g).

MS (APCI) 399 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 7.34–7.19 (5H, m), 6.66–5.68 (2H, s), 5.02–4.99 (1H,m), 4.50–4.34 (2H, m), 3.20 (1H, m), 2.73–2.68 (1H, m), 2.40 (4H, m),2.29–2.19 (2H, m), 2.05–2.00 (1H, m), 1.44–1.26 (2H, m)

Example 4[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(ethylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(ethylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

A solution of product from Example 3, step b) (0.15 g) intetrahydrofuran (10 ml) was treated with sodium ethanethiolate (0.055 g)in water (1 ml) and the reaction mixture stirred at room temperatureovernight. The solution was concentrated in vacuo and residue purifiedby chromatography (SiO₂, ethyl acetate:dichloromethane 1:1 as eluant) toafford the title compound (0.06 g).

MS (APCI) 413 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 7.34–7.20 (5H, m), 6.66 (1H, s), 5.55 (1H, s),5.05–4.96 (1H, m), 4.50–4.35 (2H, m), 3.22–2.38 (5H, m), 2.30–2.20 (2H,m), 2.05–1.95 (1H, m), 1.43–1.17 (5H, m).

Example 5[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared using the product from Example 2, stepf) and cyclopropanamine as described in Example 1, step d).

MS (APCI) 351 (M+H⁺, 100%)

NMR δH(d₆-DMSO) 9.06–9.04 (1H, m), 5.04–4.75 (3H, m), 4.49–4.42 (1H, m),4.05 (1H, m), 3.12–3.04 (3H, m), 2.37–1.98 (3H, m), 1.76–1.67 (3H, m),1.01–0.96 (3H, t), 0.87–0.67 (3H,m).

Example 6[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(3,4-dichlorophenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a)[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(propylsulphonyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared using the product from Example 5 asdescribed in Example 3, step b).

MS (APCI) 383 (M+H⁺, 100%).

b)[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(3,4-dichlorophenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared using the product from step a) and3,4-dichlorothiophenol as described in Example 3, step c) but with 60%NaH in N,N-dimethylformamide (10 ml).

MS (APCI) 453 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.22–9.20 (1H, d), 7.97–7.56 (3H, m), 4.98–4.71 (3H,m), 4.31–3.84 (2H, m), 2.92–2.89 (1H, m), 2.20–2.18 (1H, m), 1.84–1.54(4H, m), 0.88–0.87 (1H, m), 0.69–0.63 (3H,m).

Example 7[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(4-trifluoromethylphenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared using the product from Example 6, stepa) and 4-trifluoromethylthiophenol as described in Example 6, step b).

MS (APCI) 453 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.20–9.19 (1H, m), 7.90–7.79 (4H, m), 5.04–4.89 (2H,m), 4.69–4.65 (1H, m), 4.28–4.21 (1H, m), 3.83–3.78 (1H, m), 2.94–2.88(1H, m), 2.27–2.13 (1H, m), 1.82–1.80 (1H,m), 1.51 (1H, m), 0.67–0.65(3H, t).

Example 8[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

To a suspension of NaH (28 mg) in tetrahydrofuran (10 ml) was addedbutanethiol (63 mg), after 10 minutes the product from Example 3 step b)(200 mg) was added in tetrahydrofuran (1 ml). The mixture was stirredfor 12 hours before addition of saturated brine (25 ml), the organicproducts were extracted into ethyl acetate (2×25 ml), dried (MgSO₄) andconcentrated to an oil. The residue was purified by chromatography(SiO₂, methanol:dichloromethane 1:25 as eluant) to afford the titlecompound (0.087 g).

MS (APCI) 441 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.32 (1H, d), 7.29 (2H, m), 7.18 (3H, m), 5.00 (2H, m),4.75 (1H, d), 4.47 (1H, m), 4.05 (1H, m), 3.20 (1H, m), 2.80–3.00 (2H,m), 2.30 (2H, m), 1.90–20 (3H, m), 1.71 (1H, m), 1.41 (2H, m), 1.32 (2H,m), 0.81 (3H, t).

Example 9[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(3,3,3-trifluoropropylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared as described in Example 8 using theproduct from Example 3, step b) and 3,3,3-trifluoropropanethiol

MS (APCI) 481 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 7.10–7.30 (5H, m), 5.03 (2H, m), 4.76 (1H, d), 4.47(1H, m), 4.00 (1H, m), 3.05–3.20 (3H, m), 2.50 (2H, m), 2.00–2.35 (m,4H), 1.70 (1H, m), 1.45 (1H, m), 1.27 (1H, m).

Example 10[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Chlorophenyl)cyclopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a)[1R-1α,2α,3β(1R*,2S*)1-3-[7-[2-(4-Chlorophenyl)cyclopropyl]amino]-5-(propylylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The subtitle compound was prepared as described in Example 2, step g)using the product from Example 2, step f) and2-(4-chlorophenyl)cyclopropanamine (prepared as described in WO9905143).

MS (APCI) 461 (M+H⁺, 100%).

b)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Chlorophenyl)cyclopropyl]amino]-5-(propylsulphonyl)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The subtitle compound was prepared as described in Example 3, step b)using the product from Example 10, step a).

MS (APCI) 493 (M+H⁺, 100%)

c)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Chlorophenyl)cyclopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared as described in Example 8 using theproduct from step b) and butanethiol.

MS (APCI) 475 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.35 (1H, d), 7.30 (2H, d), 7.20 (3H, m), 5.03 (2H, m),4.80 (1H, m), 4.42 (1H, m), 4.05 (1H, m), 3.10–3.20 (1H, m), 2.90 (2H,m), 2.20–2.40 (1H, m), 2.00–2.19 (2H, m), 1.20–1.80 (8H, m), 0.81 (3H,t).

Example 11[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Fluorophenyl)cyclopropyl]amino]-5(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

a)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Fluorophenyl)cyclopropyl]amino]-5-(propylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The subtitle compound was prepared as described in Example 2 step g)using the product from Example 2, step f) and2-(4-fluorophenyl)cyclopropylamine (prepared as described in WO9905143).

MS (APCI) 445 (M+H⁺, 100%).

b)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Fluorophenyl)clopropyl]amino]-5-(propylsulphonyl)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The subtitle compound was prepared as described in Example 3, step b)using the product from step a).

MS (APCI) 477 (M+H⁺, 100%)

c)[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Fluorophenyl)clopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol

The title compound was prepared as described in Example 8 using theproduct from step b) and butanethiol

MS (APCI) 459 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.32 (1H, d), 7.25 (2H, m), 7.11 (2H, t), 5.10 (2H, m),4.77 (1H, d), 4.42 (1H, m), 4.08 (1H, s), 3.16 (1H, m), 2.80–3.00 (2H,m), 2.30 (1H, m), 2.17 (1H, d), 2.04 (1H, m), 1.70 (1H, m), 1.40–1.60(3H, m), 1.20–1.40 (3H, m), 0.81 (3H, t).

Example 12[1R-[1α,2β(1R*,2S*)]]-2-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

a) (trans)-2-Aminocyclopentanol

To a solution of cyclopentane oxide (5 g) in ethanol (5 ml) was added0.88 ammonia (5 ml) and the mixture heated at reflux for 9 hours. Thesolution was diluted with water (100 ml), extracted with ether (3×50ml), the organic phases were combined, washed with saturated brine (2×50ml), dried (MgSO₄) and evaporated in vacuo to afford the sub-titlecompound as an oily solid (8.1 g) that was used without furtherpurification.

b)(trans)-2-[[6-Chloro-5-nitro-2-(propylthio)pyrimidin-4-yl]amino]cyclopentanol

The sub-title compound (3.26 g) was prepared using the product from stepa) (8.1 g) and 4,6-dichloro-5-nitro-3-propylthiopyrimidine (prepared asdescribed in WO 9703084) (20 g) as described in Example 1, step a) toafford the sub-title compound.

MS (APCI) 330 (M+H⁺, 94%).

c)(trans)-2-[7-Chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

The sub-title compound (2.63 g) was prepared using the product from stepb) (3.8 g) as described in Example 1, step b).

MS (APCI) 314 (M+H⁺, 95%)

d)[(trans)-2(1R-2S)1-2-[7-1(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol(mixture of diastereomers)

The title compounds as a mixture of diastereomers (0.36 g) were preparedusing the product from step c) (0.5 g) and(1R-trans)-2-phenylcyclopropanamine,[R-(R*,R*)]-2,3-dihydroxybutanedioate (1:1) (Prepared as described by L.A. Mitscher et al, J. Med. Chem., 1986, 29, 2044) (0.45 g) andN,N-diisopropylethylamine (0.55 ml) as described in Example 1, step d).

MS (APCI) 441 (M+H⁺, 100%).

e)[1R-[1α,2β(1R*,2S*)]]-2-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

The diastereomers from step d) (0.3 g) were separated usingsupercritical fluid chromatography (Gilson SF3, Chiralpak AD column®,3000 psi, ethanol:carbon dioxide. 35:65 as solvent) to give the to givethe title compound (0.1 g) (and the[1S-[1α,2β(1S*,2R*)]]diastereomer).

MS (APCI) 441 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 0.82 (3H,t); 1.31–2.14 (11H, m); 2.82–2.94 (2H,m);3.18–3.22 (1H, m); 4.53 (1H, t); 4.79–4.86 (1H, q); 5.14–5.19 (1H,d);7.15–7.31 (5H, m); 6.33–6.16 (1H, d).

Example 13[1R-[1α,2β(1R*,2S*)]]-2-[[7-[2-(4-Methoxyphenyl)cyclopropyl]amino]-5(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

a)(1R-trans)-2-[[5-Amino-6-chloro-2-(propylthio)-4-pyrimidinyl]amino]cyclopentanol

A mixture of (1R-trans)-2-aminocyclopentanol (prepared as described inA. A. Barr et al., Can. J. Chem., 1997, 55, 4180) (3.0 g) and4,6-dichloro-3-(propylthio)pyrimidin-5-amine (6.1 g) (prepared asdescribed in EP 508687) in n-butanol (100 ml) containingN,N-diethylisopropylamine (10 ml) was heated at 100° C. for 8 hours. Thereaction mixture was evaporated to dryness and the residue taken up into2N hydrochloric acid (300 ml), washed with ether (100 ml) and thenneutralised with 0.88 ammonia solution, to afford the sub-title compound(4.0g).

MS (APCI) 303 (M+H⁺, 100%).

b)(1R-trans)-2-[7-Chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

Prepared using the product of step a) (4.0 g) by the method of Example 1step c). Purified by chromatography (SiO₂, ethyl acetate:dichloromethane1:9 as eluant) to afford the sub-title compound (4.1 g).

MS (APCI) 314 (M+H⁺, 100%)

(c)[1R-[1α,2β(1R*,2S*)]]-2-[7-[[2-(4-Methoxyphenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

Prepared using the product from step b) (0.15 g) and(1R,2S)-2-(4-methoxyphenyl)cyclopropanamine,(2R,3R)-2,3-dihydroxybutanedioate (1:1) (0.20 g) (prepared as describedin WO 9905143) by the method of Example 1, step d). Purified bychromatography (SiO₂, ethyl acetate:dichloromethane 1:9 as eluant) toafford the title compound (0.12 g).

MS (APCI) 441 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.28 (1H, d), 7.15 (2H, d), 6.84–6.86 (2H, d), 5.19(1H, d), 4.83 (1H, m), 4.55 (1H, m), 3.72 (3H, s), 3.1 1 (1H, m),2.90–2.97 (2H, m), 2.23 (1H, m), 2.11–2.23 (2H, m), 1.87 (2H, m), 1.67(2H, m), 1.57 (2H, m), 1.42 (1H, m), 1.23 (1H, m), 0.84 (3H, t).

Example 14[1R-[1α,2β(1R*,2S*)]]-2-[7-[[2-(3,4-Difluorophenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol

Prepared using the product from Example 13 step b) (0.15 g) and(1R,2S)-2-(3,4-difluorophenyl)cyclopropanamine,(2R,3R)-2,3-dihydroxybutanedioate (1:1) (0.20 g) (prepared as describedin WO 9905143) by the method of Example 13, step c). Purified bychromatography (SiO₂, ethyl acetate:dichloromethane 1:9 as eluant) toafford the title (0.12 g).

MS (APCI) 447 (M+H⁺, 100%).

NMR δH (d₆-DMSO) 9.34 (1H, d), 7.27–7.38 (2H, m), 7.07 (1H, m), 5.18(1H, d), 4.81 (1H, q), 4.51 (1H, t), 3.13–3.15 (1H, m), 2.83–2.95 (2H,m), 2.23–2.27 (1H, m), 2.07–2.17 (2H, m), 1.83–1.91 (2H, m), 1.61–1.68(2H, m), 1.41–1.57 (3H, m), 1.38–1.40 (1H, m), 0.81 (3H, t).

Pharmacological Data

The preparation for the assay of the P_(2T) (P2Y_(ADP) or P2T_(AC))receptor agonist/antagonist activity in washed human platelets for thecompounds of the invention was carried out as follows.

Human venous blood (100 ml) was divided equally between 3 tubes, eachcontaining 3.2% trisodium citrate (4 ml) as anti-coagulant. The tubeswere centrifuged for 15 minutes at 240G to obtain a platelet-rich plasma(PRP) to which 300 ng/ml prostacyclin was added to stabilize theplatelets during the washing procedure. Red cell free PRP was obtainedby centrifugation for 10 minutes at 125G followed by furthercentrifugation for 15 minutes at 640G. The supernatant was discarded andthe platelet pellet resuspended in modified, Calcium Free Tyrodesolution (10 ml) (CFT), composition: NaCl 137 mM, NaHCO₃ 11.9 mM,NaH₂PO₄ 0.4 mM, KCl 2.7 mM, MgCl₂ 1.1 mM, dextrose 5.6 mM, gassed with95% 02/5% CO₂ and maintained at 37° C. Following addition of a further300 ng/ml PGI₂, the pooled suspension was centrifuged once more for 15minutes at 640G. The supernatant was discarded and the plateletsresuspended initially in 10 ml CFT with further CFT added to adjust thefinal platelet count to 2×10⁵/ml. This final suspension was stored in a60 ml syringe at 3° C. with air excluded. To allow recovery fromPGI₂-inhibition of normal function, platelets were used in aggregationstudies no sooner than 2 hours after final resuspension.

In all studies, 3 ml aliquots of platelet suspension were added to tubescontaining CaCl₂ solution (60 μl of 50 mM solution with a finalconcentration of 1 mM). Human fibrinogen (Sigma, F 4883) and8-sulphophenyltheophylline (8-SPT which was used to block any P₁-agonistactivity of compounds) were added to give final concentrations of 0.2mg/ml (60 μl of 10 mg/ml solution of clottable protein in saline) and300 nM (10 μl of 15 mM solution in 6% glucose), respectively. Plateletsor buffer as appropriate were added in a volume of 150 μl to theindividual wells of a 96 well plate. All measurements were made intriplicate in platelets from each donor.

The agonist/antagonist potency was assessed as follows

Aggregation responses in 96 well plates were measured using the changein absorbance given by the plate reader at 660 nm. Either a Bio-TecCeres 900C or a Dynatech MRX was used as the plate reader.

The absorbance of each well in the plate was read at 660 nm to establisha baseline figure. Saline or the appropriate solution of test compoundwas added to each well in a volume of 10 μl to give a finalconcentration of 0, 0.01, 0.1, 1, 10 or 100 mM. The plate was thenshaken for 5 minutes on an orbital shaker on setting 10 and theabsorbance read at 660 nm.

Aggregation at this point was indicative of agonist activity of the testcompound. Saline or ADP (30 mM; 10 μl of 450 mM) was then added to eachwell and the plate shaken for a further 5 minutes before reading theabsorbance again at 660 nm.

Antagonist potency was estimated as a % inhibition of the control ADPresponse to obtain an IC₅₀. Compounds exemplified have pIC₅₀ values ofmore than 5.0.

1. A compound of formula (I):

a pharmaceutically acceptable salt thereof, wherein: R¹ is hydrogen orhydroxy; R² is C₁₋₆ alkyl, or C₁₋₆ haloalkyl, or phenyl optionallysubstituted by halogen or by C₁₋₆ alkyl, optionally substituted byhalogen; R³ is C₃₋₆ cycloalkyl, optionally substituted by R⁴; R⁴ ishydrogen or phenyl, optionally substituted by C₁₋₆ alkyl, halogen, orC₁₋₆ alkoxy.
 2. A compound according to claim 1 which is:

where R¹, R² and R³ are as defined in claim
 1. 3. A compound accordingto claim 2 in which R³ where R³ is

wherein R⁴ is hydrogen or phenyl, optionally substituted by C₁₋₆ alkyl,halogen, or C₁₋₆ alkoxy.
 4. A compound according to claim 1 in which R²is C₁₋₄ alkyl, or C₁₋₄ haloalkyl, or phenyl, optionally substituted byhalogen, or by C₁₋₄ alkyl, optionally substituted by halogen.
 5. Acompound according to claim 3 in which R²is C₁₋₄ alkyl, or C₁₋₄haloalkyl, or phenyl, optionally substituted by halogen, or by C₁₋₄alkyl, optionally substitued by halogen.
 6. A compound according toclaim 1, in which R³ is cyclopropyl, optionally substituted by R⁴.
 7. Acompound according to claim 4, in which R³ is cyclopropyl, optionallysubstituted by R⁴.
 8. A compound according to claim 5, in which R³ iscyclopropyl, optionally substituted by R⁴.
 9. A compound according toclaim 1, in which R⁴ is hydrogen or phenyl, optionally substituted byhalogen or C₁₋₄ alkoxy.
 10. A compound according to claim 3, in which R⁴is hydrogen or phenyl, optionally substituted by halogen or C₁₋₄ alkoxy.11. A compound according to claim 5, in which R⁴ is hydrogen or phenyl,optionally substituted by halogen or C₁₋₄ alkoxy.
 12. A compoundaccording to claim 8, in which R⁴ is hydrogen or phenyl, optionallysubstituted by halogen or C₁₋₄ alkoxy.
 13. A compound according to claim1 which is:[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α2,α3β(1R*,2S*)]]-3-[7-[[2-(4-Methylphenyl)cyclopropyl]amino-]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(methylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(ethylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(3,4-dichlorophenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-(1α,2α,3β)]-3-[7-(Cyclopropylamino)-5-(4-trifluoromethylphenylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[(2-Phenylcyclopropyl)amino]-5-(3,3,3-trifluoropropylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2α,3β(1R*,2S*)]]-3-[7-[[2-(4-Chlorophenyl)cyclopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2α,3β(1R,2S*)]]-3-[7[[2-(4-Fluorophenyl)cyclopropyl]amino]-5-(butylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]cyclopentane-1,2-diol;[1R-[1α,2β(1R*,2S*)]]-2-[7[(2-Phenylcyclopropyl)amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol;[1R-[1α,2β(1R*,2S*)]]-2-[[7-[2-(4-Methoxyphenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol;[1R-[1α,2β(1R*,2S*)]]-2-[7-[[2-(3,4-Difluorophenyl)cyclopropyl]amino]-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]cyclopentanol;or a pharmaceutically acceptable salt thereof.
 14. A pharmaceuticalcomposition comprising a compound according to claim 1 in combinationwith a pharmaceutically acceptable diluent, adjuvent or carrier.
 15. Amethod of treatment of unstable or stable angina, comprisingadministering a therapeutically effective amount of a pharmaceuticalcomposition comprising a compound according to claim 1 to a person inneed of treatment for such a condition.
 16. A method of treatment ofunstable or stable angina which comprises administering atherapeutically effective amount of a compound according to claim 1 to aperson in need of treatment for such a condition.